Sodium bis[4-hydroxy-3-[(2-hydroxy-1-naphthyl)azo]-N-(3-methoxypropyl)benzenesulphonamidato(2-)]cobaltate(1-)

Endpoint summary

Description of key information

The test item was not compatible with the in vitro skin model test system.

The test item did not show irritant properties on the skin of rabbits in vivo.

The test item gave no conclusive results in the BCOP assay.

The test item did not show irritant properties on the eyes of rabbits in vivo.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Animal Information
Two New Zealand White (Hsdlf:NZW) strain rabbits were supplied by Envigo RMS (UK) Limited, Leicestershire, UK. At the start of the study the animals weighed 2.65 or 3.13 kg and were 12 to 52 weeks old. After an acclimatization period of at least 5 days each animal was given a number unique within the study which was written with a black indelible marker pen on the inner surface of the ear and on the cage label.
Animal Care and Husbandry
The animals were individually housed in suspended cages. Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Envigo RMS (UK) Limited, Oxon, UK) was allowed throughout the study. The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 17 to 23 °C and 30 to 70% respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness.
The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

other: test material was moistened with 0.5 ml of distilled water.

Controls:

no

Amount / concentration applied:

0.5 g of the test item, moistened sufficiently with 0.5 mL of distilled water, was introduced under a 2.5 cm x 2.5 cm cotton gauze patch

Duration of treatment / exposure:

4 hours

Observation period:

7 days

Number of animals:

2

Details on study design:

On the day before the test each of a group of two rabbits was clipped free of fur from the dorsal/flank area using veterinary clippers. Only animals with a healthy intact epidermis by gross observation were selected for the study.
On the day of the test a suitable test site was selected on the back of each rabbit. At each test site a quantity of 0.5 g of the test item, moistened sufficiently with 0.5 mL of distilled water, was introduced under a 2.5 cm x 2.5 cm cotton gauze patch. The patch was secured in position with a strip of surgical adhesive tape. To prevent the animals interfering with the patches, the trunk of each rabbit was wrapped in an elasticated corset and the animals were returned to their cages for the duration of the exposure period.
Four hours after application the corset and patches were removed from each animal and any residual test item removed by gentle swabbing with cotton wool soaked in 74% industrial methylated spirits.
Immediately following removal of the patches and approximately 1, 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored according to the following scale:

Irritation parameter:

other: Erythema/Eschar Formation

Basis:

animal: 75365 Male

Time point:

other: Mean at 24, 48 and 72 hours

Score:

1.7

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

other: Erythema/Eschar Formation

Basis:

animal: 75371 male

Time point:

other: mean at 24, 48 and 72 hours

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

other: Oedema Formation

Basis:

animal: 75365 male

Time point:

other: mean of 24, 48 and 72 hours

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

other: Oedema Formation

Basis:

animal: 75371 male

Time point:

other: Mean at 24, 48 and 72 hours

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

The individual scores for erythema/eschar and edema are given in Appendix 1.
Very slight erythema and very slight edema were noted at both treated skin sites immediately after patch removal. Well defined erythema and very slight or slight edema were noted at both treated skin sites 1 hour after patch removal and at the 24 Hour observation. Well defined erythema and very slight edema were noted at one treated skin site and very slight erythema and very slight edema were noted at the other treated skin site at the 48 Hour observation. Very slight erythema, with or without very slight edema, were noted at both treated skin sites at the 72 Hour observation.
Light brown discoloration of the epidermis was noted at both treated skin sites 1 and 24 hours after patch removal.
Both treated skin sites appeared normal at the 7 Day observation.

Other effects:

Body Weight
Individual body weights and body weight change are given in Appendix 2.
Both animals showed expected gain in body weight during the study

Appendix1     Skin Irritation Scores

Individual Scores for Skin Irritation

Rabbit Number and Sex	Observation Time (following patch removal)	Erythema/Eschar Formation	Edema Formation
75365Male	Immediate	1	1
75371Male		1	1
75365Male	1 Hour	2Br	2
75371Male		2Br	1
75365Male	24 Hour	2Br	2
75371Male		2Br	1
75365Male	48 Hour	2	1
75371Male		1	1
75365Male	72 Hour	1	1
75371Male		1	0
75365Male	7 Days	0	0
75371Male		0	0

Br=  Light brown discoloration of the epidermis

Appendix 1 (continued)       Skin Irritation Scores

Mean Values after 24, 48 and 72 Hours

Rabbit Number and Sex	Number of available data points	Erythema/Eschar Formation	Edema Formation
75365Male	3	1.7	1.3
75371Male	3	1.3	0.7

Assessment According to Regulation (EC) No. 1272/2008

Evaluated Intervals	Erythema/Eschar Formation	Edema Formation
24 Hours	Not classified	Not classified
48 Hours
72 Hours

 

Appendix 2     Individual Body Weights and Body Weight Change

Rabbit Number and Sex	Individual Body Weight (kg)		Body Weight Change (kg)
	Day 0	Day 7
75365Male	3.13	3.33	0.20
75371Male	2.65	2.87	0.22

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

According to the findings in this study, the test item does not meet the criteria for classification according to Regulation (EC) No. 1272/2008 of the European Parliament and of the Council of 16 December 2008.

Executive summary:

The skin irritation potential of the test item was investigated according to a method compatible with OECD test guideline No. 404 and Method B4. 0.5 g of the test item was applied to the intact skin of two young adult New Zealand White rabbits. Scoring of skin irritation effects was performed approximately 1, 24, 48 and 72 hours and 7 days after test item application. 

The mean score was calculated separately for each animal across three scoring times (24, 48 and 72 hours after application) for erythema and edema. The individual mean scores for erythema were 1.3 or 1.7 and 0.7 or 1.3 for edema. 

The application of the test item onto

the skin resulted in well-defined erythema, very slight or slight edema and light brown discoloration of the epidermis. These effects were reversible and were no longer evident 7 days after treatment in both animals (end of the observation period). No corrosion was observed at any of the measuring intervals. No clinical signs of toxicity were observed.

Thus, the test item did not induce significant or irreversible damage to the rabbit skin.

According to the findings in this study, the test item

does not meet the criteria for classification according to Regulation (EC) No. 1272/2008 of the European Parliament and of the Council of 16 December 2008.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Two New Zealand White (Hsdlf:NZW) strain rabbits were supplied by Envigo RMS (UK) Limited, Leicestershire, UK. At the start of the study the animals weighed 2.39 or 2.53 kg and were 12 to 20 weeks old. After an acclimatization period of at least 5 days each animal was given a number unique within the study which was written with a black indelible marker pen on the inner surface of the ear and on the cage label.

The animals were individually housed in suspended cages. Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Envigo RMS (UK) Limited, Oxon, UK) was allowed throughout the study. The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 17 to 23 °C and 30 to 70% respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness.
The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study

Vehicle:

unchanged (no vehicle)

Controls:

other: The left eye remained untreated and was used for control purposes.

Amount / concentration applied:

A volume of 0.1 mL of the test item, which was found to weigh approximately 89 mg

Duration of treatment / exposure:

Up to 1 hour

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

2

Details on study design:

Immediately before the start of the test, both eyes of the provisionally selected test rabbits were examined for evidence of ocular irritation or defect with the aid of a light source from a standard ophthalmoscope. Only animals free of ocular damage were used.
Initially, a single rabbit was treated. A subcutaneous injection of buprenorphine 0.01 mg/kg was administered 60 minutes prior to test item application to provide a therapeutic level of systemic analgesia. Five minutes prior to test item application, a pre dose anesthesia of ocular anesthetic (two drops of 0.5% tetracaine hydrochloride) was applied to each eye.
A volume of 0.1 mL of the test item, which was found to weigh approximately 89 mg (as measured by gently compacting the required volume into an adapted syringe) was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test item, and then released. The left eye remained untreated and was used for control purposes. Immediately after administration of the test item, an assessment of the initial pain reaction was made according to the six point scale shown in Annex 1.
Eight hours after test item application, a subcutaneous injection of post dose analgesia, buprenorphine 0.01 mg/kg and meloxicam 0.5 mg/kg, was administered to provide a continued therapeutic level of systemic analgesia. The treated animal was checked for signs of pain and suffering approximately 12 hours later. No further analgesia was required.
After consideration of the ocular responses produced in the first treated animal, a second animal was similarly treated.
Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation (Draize, J.H, 1977) given in Annex 2.
Any other ocular effects were also noted. Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.
Any clinical signs of toxicity, if present, were also recorded.
Individual body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

Irritation parameter:

cornea opacity score

Basis:

animal: 75330 Female

Time point:

other: Mean 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: No effects observed

Irritation parameter:

cornea opacity score

Basis:

animal: 75321 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: No effects observed

Irritation parameter:

iris score

Basis:

animal: 75330 Female

Time point:

other: Mean 24, 48 and 72 hours

Score:

0

Max. score:

2

Reversibility:

other: No effects observed

Irritation parameter:

iris score

Basis:

animal: 75321 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0

Max. score:

2

Reversibility:

other: No effects observed

Irritation parameter:

other: redness

Basis:

animal: 75330 Female

Time point:

other: Mean 24, 48 and 72 hours

Score:

0.7

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

other: redness

Basis:

animal: 75321 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0.7

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

chemosis score

Basis:

animal: 75330 Female

Time point:

other: Mean 24, 48 and 72

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

chemosis score

Basis:

animal: 75321 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritant / corrosive response data:

Individual and mean scores for ocular irritation are given in Appendix 1.
Red colored staining of the fur around the treated eye was noted in both animals at all observations.
No corneal or iridial effects were noted during the study.
Moderate conjunctival irritation was noted in both treated eyes 1 hour after treatment with minimal conjunctival irritation noted at the 24 and 48-Hour observations.
Both treated eyes appeared normal at the 72-Hour observation.
No corrosive effects were noted during the study. The test item did not induce significant or irreversible damage to the rabbit eye.

Other effects:

Body weight
Individual body weights and body weight change are given in Appendix 2.
Both animals showed expected gain in bodyweight during the study.

Appendix 1     Eye Irritation Scores

Individual Scores for Eye Irritation

Rabbit Number and Sex	IPR	Evaluation interval	Corneal Opacity	Area of Corneal Opacity	Iris	Conjunctivae
						Redness	Chemosis	Discharge
75330Female	0	1 Hour	0	0	0	2	1	1Sf
75321Male	0		0	0	0	2	1	2Sf
75330Female		24 Hour	0	0	0	1	1	0Sf
75321Male			0	0	0	1	1	1Sf
75330Female		48 Hour	0	0	0	1	0	0Sf
75321Male			0	0	0	1	0	0Sf
75330Female		72 Hour	0	0	0	0	0	0Sf
75321Male			0	0	0	0	0	0Sf

IPR=Initial pain reaction

Sf =       Red colored staining of the fur around the treated eye

Mean Values after 24, 48 and 72 Hours

Rabbit Number and Sex	Number of available data points	Corneal Opacity	Iris	Conjunctivae
				Redness	Chemosis
75330Female	3	0.0	0.0	0.7	0.3
75321Male	3	0.0	0.0	0.7	0.3

Assessment According to Regulation (EC) No. 1272/2008

Evaluated Intervals	Corneal Opacity	Iris	Conjunctivae
			Redness	Chemosis
24 Hours	Not classified	Not classified	Not classified	Not classified
48 Hours
72 Hours

Appendix 2     Individual Body Weights and Body Weight Change

Rabbit Number and Sex	Individual Body Weight (kg)		Body Weight Change (kg)
	Day 0	Day 3
75330Female	2.53	2.61	0.08
75321Male	2.39	2.40	0.01

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test item did not induce significant or irreversible damage to the rabbit eye.
According to the findings in this study, the test item does not meet the criteria for classification according to Regulation (EC) No. 1272/2008 of the European Parliament and of the Council of 16 December 2008.

Executive summary:

The primary eye irritation potential of the test item was investigated according to a method compatible with OECD test guideline No. 405 and Method B5. The test item was applied by instillation of 0.1 mL into the right eye of each of two young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours after test item instillation. 

The mean score was calculated separately for each animal across three scoring times (24, 48 and 72 hours after instillation) for corneal opacity, iritis, redness and chemosis of the conjunctivae. The individual mean scores for corneal opacity and iritis was 0.0 for both animals. The individual mean scores for the conjunctivae were 0.7 for reddening and 0.3 for chemosis for both animals. 

The instillation of the test item into the eye resulted in conjunctival irritation. These effects were reversible and were no longer evident 72 hours after treatment in both animals (end of the observation period). No abnormal findings were observed in the cornea of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. No clinical signs of toxicity were observed.

Thus, the test item did not induce significant or irreversible damage to the rabbit eye.

According to the findings in this study, Savinyl Red 3BLS does not meet the criteria for classification according to Regulation (EC) No. 1272/2008 of the European Parliament and of the Council of 16 December 2008.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

No classification

No adverse effects were detected.